4-chloro-adrenosterone



United States Pat n 2,953,582 I 4-CI-ILORO-ADRENOSTERONE Bruno Camerino, Milan, Italy, assignor to Societa Farmaceutici Italia, a corporation of Italy No Drawing. Filed Oct. 26, 1956, Ser. No. 618,441 Claims priority, application Italy Apr. 23, 1956 1 Claim. (Cl. 260397.3)

This invention relates to a new class of steroids characterized by high anabolic activity coupled with low androgen activity. a

The herein claimed compounds are prepared by a modification of the process disclosed'in the copending application of July 19, 1956, SerialNo; 598,754,-nw abandoned, entitled New Steroid Hormone Derivatives Substituted in the 4-Position and Method of Preparing Same of which this application is a continuation-in-part. The compounds disclosed in said copending application are prepared by reacting 4,5-epoxy-3-keto-steroids of the general formula v I wherein R may represent =0, (H)OI-I and (CH )OH, and R represents H =0 and (H) OH with mineral acid in an organic solvent.

The compounds. obtained according to this invention have the general formula L j;

R represents F, Cl and R represents acyl and alkoxy.

According to the aforementioned, copending patent application, steroids wherein R represents F or Cl are prepared from only the fi-form of 4,5-epoxides.

We have now found that steroids of this type can be also obtained from the u-form of 4,5-epoxides upon treating with halogenhydric acids in chloroform *sol ution or in a chloroform solution containing absolute ethanol. Moreover, if R'" is to become Cl, it is suflicient to heat the epoxide with a chloroform solutionofpyri- 1 dinium chloride. y

' According to the present invention, it is therefore possible to prepare 4-chloro and the 4-fluoro-steroids directly from a mixture of 4,5;8- and 4,5a-epoxides obtained by medicine. Anabolic as wellas androgen activity are ice increase in yield, coupled with a simplification of the operations. 7 I

The compounds claimed in the present invention display strong anabolic activity on proteins, while beingv devoid of any'substantial androgen activity. Therefore, they are of great importance .in human and veterinary always jointly present in steroids that have been hitherto used in an'abolitic therapy. Now we are able to provide products that do not produce any substantial androgen- .7

eff ct. a

treating a 3-keto-A -steroid with alkaline hydrogen per; 0

oxide. Consequently, the. process-resultsin a substantial Therefore, it islpossible-to take advantageof the a'nja-H bolic activity of these compounds without stimulating the sexual activity and-this property -is particularly useful for the treatment of decay, oseoporosis emaciation, convalescences, premature" newborns, underdevelopment,

senility. Y

In addition, the products of the present invention also exhibit the other properties of previously known anabolic substances on the basis of which they have beenused in treating certain ovary malfunctions, etc;

The following examples illustrate the present inven-.

tion without limiting its scope. i

EXAMPLE 1 4-chl0r0-test0sterone V 7 15 kg of, a mixture of.45,5-epoxy-etiocholane-17 3-01-1 3-one and 4a,5-epoxy-androstane-17B-ol-3-one, dissolved,

-The' residue is crystallized from benzene oraqueous methanol. 9 g. of needle-shaped crystals, M.P. 186-188 1 C.', are obtained. Upon concentrating the mother-liquor,

3.2. g. of a product having a M.P. of -184" C, are

recovered. I

A max. 256 m =13,150.

EXAMPLE 2 4-chlor0-test0ster0ne I Example 1 is'repeated, except that the chloroform is substituted by methylene chloride containing 3% methanol..

' EXAMPLE 3 4-chloro-testosterone Exarnple 1 is repeated except that the saturation with:

gaseous HCl is carried out at 10 C. for 2 hours.

EX AM PLE 4 4-chlor0-testostemite-acetate 3 no go 5 g. 45,5-epoxy-etiocholane-17fi-o1-3-one-acetate, M.P.

156 C., are dissolved in 125 cc. anhydrous chloroform and treated with a gaseous HCl stream at room temperature.

dark and turbid. After 2 hours the organic solution is Washed first with a sodium bicarbonate solution and then,

with water, and is finally evaporated to dryness whereby the drying is Completed under vacuum.

The residue is taken up with 400 cc. hot methanol, and the solution is cooled and filtrated. 3.3 g. 4-chlorotestosterone-acetate are obtained, M.P. 228-230 C., A max. 255 mu, e=13,300, [a] =+118 i4 (in chloroform).

0.5 g. of a product having a M.P. of 215-220 C. is recovered upon concentrating the filtrate.

EXAMPLE 5 4-chloro-testosterone-acetate 4 g. 4,6,5-epoxy-etioholane 1713 o1 3 one acetate, M.P. 156 C., are refluxed with 50 cc. of a 1.2/N solution of pyridinium chloride in chloroform for 5 hours. The organic solution is washed with diluted hydrochloric acid, sodium bicarbonate and water and is finally evaporated to dryness.

The crystalline residue, M.P. 220228 C., consists of almost pure 4-chloro-testosterone-acetate. After a recrystallization the product melts at 228-230 C.

EXAMPLE 6 4-chloro-test0ster0ne-acetate 2 g. 4a,5-epoxy-androstane-175-ol-3-one-acetate M.P. 170172 C., dissolved in 50 cc. anhydrous chloroform, are treated with anhydrous HCl for 2 hours at room temperature. The solution becomes pale-yellow without any further darkening. The solution is neutralized by washing and concentrated. The residue obtained upon evaporation is recrystallized from methanol and 1.5 g. 4-chloro-testosterone-acetate, M.P. 226-228 C., are obtained.

EXAMPLE 7 4-chloro-tesl0ster0ne-acetate 1 g. 4u,5-epoxy-androstane-17fl-ol-3-one-acetate are boiled for 8 hours in 15 cc. of a 1.2/N solution of pyridinium chloride in coloroform.

The resulting solution is washed with dil. hydrochloric acid, diluted alkali, water and is finally reduced to dryness. An almost quantitative yield of 4-ch1oro-testosterone-acetate is obtained.

EXAMPLE 8 4-chl0ro-test0sterone-acetate 1 g. 4-chloro-testosterone are acetylated with 1 cc. acetic anhydride and 5 cc. pyridine at room temperature for 16 hours. Ice is added to the solution, and the precipitate is filtered off and recrystallized from chloroform-ethanol; 1 g. 4-chlorotestosterone-acetate, M.P. 228- 230 C., is obtained.

EXAMPLE 9 4-chloro-testosterone-propionate O C 0 013a CH3 The solution becomes yellow-green and subsequently water.

4-chloro-testosterone-emisuccinate V (|)COCH:4OHCOOH clj 0.7 g. 4-chloro-testosterone, M.P. 188 C., are treated with 7 cc. pyridine and 1.2 g. succinic anhydride at 70 C. for 5 hours. After pouring the solution into ice-cold Water, extracting with benzene and treating the extract with diluted hydrochloric acid, washing with' water and drying by evaporation, the residue is crystallized from aqueous methanol. [a] =1l6 i4, are obtained.

EXAMPLE 11 4-chloro-test0sterone-pa'lmitate 1 g. 4-chloro-testosterone, dissolved in 10 cc. benzene and 3 cc. pyridine, are treated with 1.3 g. palmityl chlo ride at --10 C. and left standing at room temperature for 12 hours. The solution is then diluted with benzene, and washed with diluted HCl, diluted NaOH and The benzene is evaporated and the residue is recrystallized from methanol. 0.8 g. 4-chloro-testosterone-palmitate, M.P. 56 C., are obtained.

EXAMPLE 12 4-chl0r0-17a-methyl-test0sterone 3 g. 4a,5 epoxy 17'- methyl-androstane-17fl-ol-3-one,

M.P. -82 C., [a]j 12 :4, dissolved in cc. chloroform and'9 cc. ethanol, are treated with anhydrous 'HCl until the solution is saturated. After standing for 15 min., the solution is washed with Water and dried by distilling olf the solvent; The residue-is chromatographically fractionated on 90 g. Florisil and the fractions eluted with ether are recrystallized from ether. 2.2g.

4-ch'loro-17a-methyl-testosterone are obtained, M.P. 1'48 C., A max. 256 mu, e=14.000. I

0.4 g. of crystals, M.P. 203204 C.,-

. 4-chloro-A -artdros ten e -3fl 7-di0ne 7 z 2 g. 4-ch1oro-testo'sterone dissolvedfin 20;. cc. pyridine are treated, at roomtemperature, with-a suspension of 1 g .CrO in 20 cc. pyridine. After 241 hours thereaction mixture is diluted with-benzene, washed with diluted hydrochloric acid, diluted NaOH and water and is finally dried. I

The solvent is evaporated and the residue is crystallized from aqueous methanol. 0.7 g. 4-chloro-A -andro stene-3,;17-dione are obtained, M.P. 175 C., lul =+21s :4".

' EXAMPLE 14 4-chl oro-ad renosterohe 12 g. 4-chloro testosterone, dissolved in 120 cc. pure methanol, are cooled to C. and treated for 2 hours with 0.5 g. NaBH at +5 C. After a short time, a voluminous crystalline precipitate appears in the solution. This precipitate is extracted with methylene chloride and washedwi-th diluted hydrochloric acid and water. 1.7 g, residue, M.P. about 140 C., are obtained. After recrystallization from methanol, the melting point ofzthe product is 252 C.; [a] =+114 i4. No ultraviolet absorption between 220 and 300 mp.

'By acetylati on withamixture of acetic anhydride and pyridine, the 4-chloro-A -androstene-3j3-l7fi-diol-diacetate is obtained which, when recrystallized from methanol, melts at 150 C., [a] =+56 :4.

1 g. 4,5p-epoxy-etiocholane-17e-ol-3-one-acetate, dissolved in 30 cc. chloroform and 30-00. absolute ethanol, are treated with anhydrous HF for 1 hour. A 2 N NaOH solution is then added until the solution is only slightly. acid:. The' precipitate isseparated,- washed-with H O, dried and thesolvent contained therein is distilled-- off. 7 Upon recrystallization firom ether, 190 mg. 4-fluoro-- testosterone-acetate are obtained; M.P.. 178- 180" C., A

Proceeding as in Example 16, but starting with the 4fi,5-cpoxyetiocho1ane-17fi-ol-3-one-propionate, 4-fluorotestosterone-propionate is obtained by recrystallization from petroleum, M.P. 128-130" C., A max. 243 11111., 6 12.300.

7 g fEXAM PLE 18, i I

- 4 chloro-tes rostrongceiate-bxirrie i 'OOOCHa 1 g. 4-chloro-testosterone-acetate are refluxed for 2 hours with 3 g. hydroxylamine hydrochloride, 250 cc. absolute ethanol, 1 cc. pyridine and 5 cc. acetic acid. The solution is then concentrated and the precipitate is recrystallized from ethyl alcohol. 07. g. of the oxime are'obt'ained, M.P..20 t-205 C. (decomposition).

EXAMPLE 19 4- chloro-testosterone-acetate-thiosamicarbazone Hm SHNN hours with 0.6 g. thiosemicarbazide and cc. ethanol.

The solution is concentrated andthe precipitate is filtered ofl and recrystallized trom ethanol, 0.6 g. thiosemicarbazone are obtained, M.P. 197-204'C. (decomposition).

EXAMPLE 2O 4-chloro-testosterone-trim ethylacetate O C QG (C1193 Ni guy i EXAMPLE A 2 1 4-chl0r0-testosterone-pheny lprqpionate O O O CHzcHaco s 2 g. 4-chloro-testosteroneare esterified with 2 g. phenylpropionyl acid chloride as described in Example 20. 1.7 g. of the phenylpropionate, M.P. 145 C., are obtained.

4-ch loro-testosterone-enanthate O CO.(CH2) sCHa 1 g. 4-chloro-testosterone dissolved in cc. pyridine are heated for 5 hours'to 90 C. with 2 g. enanthic anhydride. The solution is diluted with water, and extracted with benzene. The benzene extract iswashed with diluted hydrochloric acid, diluted NaOH solution and water until neutral. The solvent is evaporated and the oily residue is purified chromatographically on silica gel. The fraction eluted with benzene-ether produces 1.1 g. of the enanthate, M.P. 59 C.

EXAMPLE 23 Pharmacological activity of the 4-chl0r0-test0steroneacetate 4-chloro-testosterone-acetate, injected into impuberal rats castrated according to the I lershberger et method 8 (Proc. Soc. Exp. Biol. andiMedl 83, 175 (1953)), shows high anabolic actiyityreoupled with poor. androgen activity.

Thus, a daily dose of 500 'y of the afore-mentioned 3 compound increases the weight of the levator ani muscle from 5 to 42 mg. while 500 'y testosterone propionate increase the weight of this muscle to 28.5 mg. Under similar condition 4-ch1or0-testosterone-acetate increases the prostate weight from 6.7 to 48 mg. while Tthe testosterone-propionate increases this weight to mg. The seminal bladders increase. from 4.1 mg. to 36.5 mg. as a result of the actionof the 4-chloro-testosterone-acetate and to 96 mg. as a result of the action of the testosterone-propionate.

Consequently, the ratio between the anabolic and the androgen activity, calculated according to Hershberger et al., is 0.88 for 4-chloro-testosterone-acetate and- 0.28

for testosterone-propionate.

4-chloro-testosterone-acetate is still'active in doses as little as 250 'y and 100 7 per day.

The toxicity of 4-chloro-ttestosterone-acetate is low: a dose of 1.5 g./kg. administered by one subcutaneous injection of the aqueous solution is readily tolerated by rats. Y I

Doses of 50 mg./kg.-a dayfor -1-0 days are also well tolerated by rats while a dose 10 as great may be considered the maximum dose tolerated upon continuous administration.

EXAMPLE 24 Pharmacological activity of other derivatives The 4 chloro testosterone propionate increases the weight of the levator ani muscle from 5 to 27 mg;

The 4-hydroxy-testosterone-17-acetate from 5 to 20.5 mg;

The 4-chloro-testosterone from 5 to 19 mg.;

The 4-ch1oro-l7u-methyl-testosterone from 9.8 to 24.5

The 4-fiuoro-testosterone-propionate from 7 .9-to 20mg.

The 4 chloro -testosterone -propionate increases the weight of the prostate from 6.7 to 43 mg. (ratio between I anabolic and androgen activity 0.61); the 4-hydroxytestosterone-l7-acetate from 6.7 to 32 mg. (ratio as above mentioned, 0.61)

The chloro-testosterone from6.7 to 31.5 as above-mentioned, 0.57),

The 4-chloro-17a-methyl-testosterone from 4.2 to 48.7 mg. (ratio, as afore-rnentioned, 0.35);

The 4-fluoro-testQsterone-propionate iron; 5.2 to 39.2

1 mg. (ratio, as aforementioned, 0.35).

I claim:

4-chloro-adrenosterone.

References Cited in the file of this patent UNITED STATES PATENTS 2,845,381 Tindall July 29, 1958 2,933,510 Julian et a1 Apr. 19, 1960 OTHER REFERENCES Kochakian et al.: J. Biol. Chem. vol. 122 (1938), pages 433-438 (page 433 necessary). 7

Cavallini et al.: Boll., Soc. Ital. Biol. -Sper., vol. 27

(1951), pages 629 -30; abstracted in Chem. Absh, vol.

Bremer: Congress Handbook, XIVth Internet. Cough,

Pure and Applied Chemistry (Zurich, Switz.': Berihthaus Zurich, 1955 pages 162 and 163,

Kirk et al.: I..Chem. Soc, (March 1956 pages 627 Kirk et al.:

'Camerino et al.:,I. Am. Chem. -Soc.,vol. 78 -('Iu1y.;20,

1.9.5.6), pages 3540 and 3541.

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